Methanobacterium Transduction Medium

[所属分类:培养基配方] [发布时间:2021-5-24] [发布人:网站管理员2] [阅读次数:] [返回]

Methanobacterium Transduction Medium

山东拓普生物工程有限公司 培养基配方 http://www.topbiol.com

Composition per liter:

Basal salts solution ................................................................100.0mL
Na 2 CO 3 solution ......................................................................24.0mL
Trace elements solution...........................................................10.0mL
L -Cysteine/Na 2 S solution.........................................................10.0mL
Basal Salts Solution:
Composition per liter:
KH 2 PO 4 .......................................................................................68.0g
NH 4 Cl .........................................................................................21.2g
Resazurin .................................................................................10.0mg
Preparation of Basal Salts Solution:  Add components to dis-
tilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N 2 .
Trace Elements Solution:
Composition per liter:
Nitrilotriacetic acid.....................................................................9.55g
MgCl 2 ·7H 2 O ...............................................................................4.06g
FeCl 2 ·7H 2 O.................................................................................0.98g
NaWO 4 ·6H 2 O ...........................................................................0.264g
NiCl 2 ·6H 2 O............................................................................... 0.118g
Na 2 Se 2 O 3 ......................................................................................0.1g
NaMoO 4 ·2H 2 O..........................................................................0.024g
CoCl 2 ·6H 2 O..............................................................................0.023g
Preparation of Trace Elements Solution: Add nitrilotriacetic
acid to 500.0mL of distilled/deionized water. Adjust pH to 7.0 with
KOH. Add remaining components one at a time. Add distilled/deion-
ized water to 1.0L. Adjust pH to 7.0. Sparge with 100% N 2 .
Na 2 CO 3 Solution:
Composition per liter:
Na 2 CO 3 .......................................................................................10.6g
Preparation of Na 2 CO 3 Solution:  Add Na 2 CO 3 to distilled/de-
ionized water and bring volume to 100.0mL. Mix thoroughly. Sparge

with 80% N 2 + 20% CO 2 .

L -Cysteine/Na 2 S Solution:
Composition per 50.0mL:
L -Cysteine·HCl..............................................................................5.0g
Na 2 S·9H 2 O....................................................................................5.0g
Preparation of  L -Cysteine/Na 2 S Solution:  Gently heat and
bring 50.0mL of distilled/deionized water to boiling. Add  L -
cysteine·HCl and Na 2 S·9H 2 O. Mix thoroughly. Cool while sparging
with 100% N 2 . Bring volume to 50.0mL with distilled/deionized water.
Sparge with 100% N 2 . Adjust pH to 10.0 with 3N NaOH.
Preparation of Medium: Prepare and dispense medium under 80%
N 2 + 20% CO 2 . Add components to distilled/deionized water and bring
volume to 1.0L. Mix thoroughly. Sparge with 80% N 2 + 20% CO 2 . An-
aerobically distribute into tubes or bottles. Autoclave for 15 min at 15
psi pressure–121°C. Prior to inoculation, pressurize to 200kPa with an
atmosphere of 80% H 2 + 20% CO 2 .
Use: For the cultivation of Methanobacterium species for transduc-
tion.